IgG2 DISULFIDE ISOFORM SEPARATION
AssigneeAmgen Inc.
InventorsThomas M. Dillon, Yi-Te Chou, Yautyan Chen, Pavel Bondarenko, Jed J. Wiltzius, Diana Woehle
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Abstract
Methods for producing an IgG2 antibody preparation enriched for one of several IgG2 structural isoforms, differing by disulfide connectivity in the hinge region of the antibody, are disclosed.
Claims22
A method of producing an IgG2 antibody preparation enriched for one of several IgG2 structural isoforms which differ by disulfide connectivity in the hinge region of the antibody, comprising
- (A) contacting a solution containing a recombinantly-produced IgG2 antibody with a first matrix selected from the group consisting of a strong cation exchange (SCX) matrix, an anti-human IgG2 affinity matrix and a Protein L matrix, and
- (B) eluting two or more first elution fractions from the first matrix,
- wherein the IgG2 antibody solution to be subjected to the method elutes off the first matrix as two or more separate forms corresponding to two or more IgG2 structural isoforms, and at least one of the two or more first elution fractions is enriched for at least one of the IgG2 structural isoforms which differ by disulfide connectivity in the hinge region.
The method of claim 14, wherein the pH gradient is varied from a pH of between 7 and 7.5 to a pH between 2.5 and 3.
A method of producing an IgG2 antibody preparation enriched for one of several IgG2 structural isoforms which differ by disulfide connectivity in the hinge region of the antibody, comprising
- subjecting a preparation comprising an IgG2 antibody having a kappa light chain of the VκI, VκIII, or VκIV subclasses to a Protein L resin,
- wherein said IgG2 antibody in said preparation elutes from said Protein L resin as two or more structural IgG2 isoforms.
Dependencies are inferred from the claim text.